Aureochrome 1 illuminated: structural changes of a transcription factor probed by molecular spectroscopy

PLoS One. 2014 Jul 24;9(7):e103307. doi: 10.1371/journal.pone.0103307. eCollection 2014.

Abstract

Aureochrome 1 from Vaucheria frigida is a recently identified blue-light receptor that acts as a transcription factor. The protein comprises a photosensitive light-, oxygen- and voltage-sensitive (LOV) domain and a basic zipper (bZIP) domain that binds DNA rendering aureochrome 1 a prospective optogenetic tool. Here, we studied the photoreaction of full-length aureochrome 1 by molecular spectroscopy. The kinetics of the decay of the red-shifted triplet state and the blue-shifted signaling state were determined by time-resolved UV/Vis spectroscopy. It is shown that the presence of the bZIP domain further prolongs the lifetime of the LOV390 signaling state in comparison to the isolated LOV domain whereas bound DNA does not influence the photocycle kinetics. The light-dark Fourier transform infrared (FTIR) difference spectrum shows the characteristic features of the flavin mononucleotide chromophore except that the S-H stretching vibration of cysteine 254, which is involved in the formation of the thio-adduct state, is significantly shifted to lower frequencies compared to other LOV domains. The presence of the target DNA influences the light-induced FTIR difference spectrum of aureochrome 1. Vibrational bands that can be assigned to arginine and lysine side chains as well to the phosphate backbone, indicate crucial changes in interactions between transcription factor and DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algal Proteins / chemistry*
  • Algal Proteins / metabolism
  • Binding Sites
  • Cryptochromes / chemistry*
  • Cryptochromes / metabolism
  • DNA / metabolism*
  • Kinetics
  • Models, Molecular
  • Protein Structure, Secondary
  • Spectroscopy, Fourier Transform Infrared
  • Structural Homology, Protein
  • Transcription Factors / chemistry*
  • Transcription Factors / metabolism

Substances

  • Algal Proteins
  • Cryptochromes
  • Transcription Factors
  • DNA

Grants and funding

This work was funded by the Deutsche Forschungsgemeinschaft (DFG) within the Forschergruppe 1279 “Protein-based photoswitches as optogenetic tools” given to IS and JH. www.dfg.de. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.